Generating Stronger T cells for Adoptive Immunotherapy

Abstract

Event Abstract Back to Event Generating Stronger T cells for Adoptive Immunotherapy Connie P. Duong1, 2*, Jennifer A. Westwood1, Carmen S. Yong1, 3, Christel Devaud1, Liza B. John1, Phillip K. Darcy1, 3, 4 and Michael H. Kershaw1, 3, 4 1 Peter MacCallum Cancer Centre, Cancer Immunology Program, Australia 2 University of Melbourne, Department of Pathology, Australia 3 University of Melbourne, Sir Peter MacCallum Department of Oncology, Australia 4 Monash University, Department of Immunology, Australia Adoptive immunotherapy mediates effective regression of advanced metastatic melanoma in patients. To target other cancers, we engineered T cells to express chimeric antigen receptors (CARs) empowering them with anti-tumour activity, specifically to the tumor-associated antigen, erbB2. Our current CAR, utilising CD28-CD3ζ signaling domains, although promising, mediates lower activity compared to endogenous TCR responses against foreign antigens. Therefore, we hypothesized that incorporating multiple alternate signaling domains into CARs could result in significantly enhanced anti-tumor responses. To investigate this, we generated a library of approximately 30,000 CARs that incorporated 14 different signaling molecules randomly ligated together directionally and in-frame flanked by SfiI sites and inserted into an anti-erbB2 CAR retroviral vector. The Jurkat T cell line was then retrovirally transduced to express the library and screened for increased CD69 expression or IL-2 secretion in response to antigen specific stimulation. Genomic DNA of selected clones was then extracted, sequenced and cloned into the pSAMEN vector to transduce human peripheral blood mononuclear cells. A diverse range of CARs containing different signaling chains were identified. A particular CAR, containing the tripartite signaling domains DAP10-CD3ζ-CD27 endowed human T cells with the ability to mediate significantly enhanced killing of erbB2+ tumor cells in vitro compared to the CD28-CD3ζ CAR (p<0.05). Strikingly, adoptive transfer of T cells expressing this novel receptor induced significantly increased anti-tumor activity in vivo (p<0.01). In conclusion, it is anticipated that the generation of CARs with novel combinations of signaling domains will lead to stronger T cells for the treatment of cancer. Keywords: immunotherapy of cancer, Chimeric Antigen Receptor, DNA library, Jurkat Cells, genetic modification, CD3 zeta, CD28, DAP10 Conference: 15th International Congress of Immunology (ICI), Milan, Italy, 22 Aug - 27 Aug, 2013. Presentation Type: Abstract Topic: Translational immunology and immune intervention Citation: Duong CP, Westwood JA, Yong CS, Devaud C, John LB, Darcy PK and Kershaw MH (2013). Generating Stronger T cells for Adoptive Immunotherapy. Front. Immunol. Conference Abstract: 15th International Congress of Immunology (ICI). doi: 10.3389/conf.fimmu.2013.02.01120 Copyright: The abstracts in this collection have not been subject to any Frontiers peer review or checks, and are not endorsed by Frontiers. They are made available through the Frontiers publishing platform as a service to conference organizers and presenters. The copyright in the individual abstracts is owned by the author of each abstract or his/her employer unless otherwise stated. Each abstract, as well as the collection of abstracts, are published under a Creative Commons CC-BY 4.0 (attribution) licence (https://creativecommons.org/licenses/by/4.0/) and may thus be reproduced, translated, adapted and be the subject of derivative works provided the authors and Frontiers are attributed. For Frontiers’ terms and conditions please see https://www.frontiersin.org/legal/terms-and-conditions. Received: 25 Jul 2013; Published Online: 22 Aug 2013. * Correspondence: Ms. Connie P Duong, Peter MacCallum Cancer Centre, Cancer Immunology Program, Melbourne, Australia, connieduong.cd@gmail.com Login Required This action requires you to be registered with Frontiers and logged in. To register or login click here. Abstract Info Abstract The Authors in Frontiers Connie P Duong Jennifer A Westwood Carmen S Yong Christel Devaud Liza B John Phillip K Darcy Michael H Kershaw Google Connie P Duong Jennifer A Westwood Carmen S Yong Christel Devaud Liza B John Phillip K Darcy Michael H Kershaw Google Scholar Connie P Duong Jennifer A Westwood Carmen S Yong Christel Devaud Liza B John Phillip K Darcy Michael H Kershaw PubMed Connie P Duong Jennifer A Westwood Carmen S Yong Christel Devaud Liza B John Phillip K Darcy Michael H Kershaw Related Article in Frontiers Google Scholar PubMed Abstract Close Back to top Javascript is disabled. Please enable Javascript in your browser settings in order to see all the content on this page.